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5) is performed on the emission spectrum from a reference solution with known concentration of fluorophore and the emission spectrum from a suspension of microspheres with immobilized FITC. It is widely accepted that describing a measured x-ray peak by a Gaussian distribution is a sufficient approximation to derive accurate x-ray intensities from an x-ray emission spectrum measured with an EDS. It has been found that emission spectrum matching between a reference solution and an analyte and normalization by the corresponding extinction coefficient are required for quantifying fluorescence signals using flow cytometers. |
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